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Integrating Light and TEM Information with F-TEM images

Published online by Cambridge University Press:  14 March 2018

P.A. Sims ,
C.A. Lockwood  and
J.D. Hardin
P.A. Sims*
Affiliation:
Zoology Department, University of Wisconsin
C.A. Lockwood
Affiliation:
Zoology Department, University of Wisconsin
J.D. Hardin
Affiliation:
Zoology Department, University of Wisconsin
*
pasims@wisc.edu

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Fluorescent fusion proteins are widely used to visualize the localization of proteins in worms, fish, flies and tissue culture cells. We have used two different methods that use high pressure freezing (HPF) combined with correlative light microscopy (LM) and TEM. The first method uses fluorescence from live organisms immobilized in agarose followed by HPF and standard freeze substitution in dry solvent with osmium. This pre-embedding method optimizes ultrastructural preservation. A second, post-embedding method preserves fluorescence and immunoreactivity from embedded and polymerized thin sections. Here we describe post-embedding fluorescent integrated TEM images (F-TEM).

Type
Research Article
Information
Microscopy Today , Volume 13 , Issue 5 , September 2005 , pp. 16 - 19
Copyright
Copyright © Microscopy Society of America 2005

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